Tecan Journal Edition 03/2007

Tecan Journal

Edition 3 / 2007

Custom-made for you page 9

Painting a picture of gene expression throughout the mouse brain pages 10 – 12

Processing 15 million aliquots to support future medical research pages 14 – 15

Cover shows Dennis von Ballmoos scanning samples for the Freedom EVO® Clinical 75 NAT.

C E O WE LCOME

There’s more to Tecan than meets the eye… …as the audience at our recent presentation at the AACC annual meeting in San Diego soon discovered. The presentation focused on Tecan’s wealth of experience in creating innovative solutions for the research sector, and also on our less well-known, but equally important, track record as an OEM provider of instruments to the world’s leading diagnostic companies. Together, these qualities serve us well in providing automation solutions for highly regulated laboratories.

Welcome to this edition of the Tecan Journal, where you’ll be able to read about just a few of these innovative solutions, and can hear all the latest news from happy customers and dedicated Tecan personnel alike. We have great news of an exciting partnership with the Monash Antibody Technologies Facility (MATF) in Australia to develop one of the largest, most sophisticated monoclonal antibody production systems in the world. We are delighted to be involved in this project in such a fast-moving and ground-breaking area of science.

Tecan Journal 3/2007

A customer in the UK explains just how easy it is to process 15 million samples with Tecan’s Freedom EVO® liquid handling workstations for a fascinating long-term patient study, and the Texas-based company Family Tree DNA describes how it manages, processes and stores samples for the Genographic Project using both REMP and Tecan instruments. There’s news too of our hugely successful, multi-functional microplate reader, the Infinite™ M200, being put through its paces by scientists at the cosmetics company Wella, plus lots more in what we hope you will find yet another informative and interesting issue of our Tecan Journal.

We hope you enjoy this issue, Thomas Bachmann, CEO

CONTE N TS CEO welcome Thomas Bachmann, CEO, welcomes you to this issue of the Tecan Journal. page 2 Product news: Tecan’s new QuadChamber™ is validated for use with Agilent’s 4-pack Microarrays Fully automated processing of four different microarrays on one slide. page 4

Quality and quantity control for pooling of clinical samples Integrated solution brings a new level of safety and quality assurance to blood pooling laboratories. page 7

Processing 15 million aliquots to support future medical research A fleet of Freedom EVO® workstations at the UK Biobank processes thousands of blood samples daily. pages 16-17 High throughput screening (HTS) systems at Boehringer Ingelheim Customized, automated high throughput systems designed by Tecan are helping Boehringer Ingelheim’s latest screening projects. pages 18-19 Clever use of color Wella uses the Infinite™ M200 for cytotoxicity testing of colored compounds. pages 20-21

Enhance your crystallography with the new CrysScreen™ 2.0 software An upgraded version of the software offers a number of user-friendly improvements in the management of protein crystallography trials. page 5

The 6th International BioForum & BioExpo Japan Tecan shows latest cell-based assay products in Japan. page 8 Custom-made for you Customized systems designed by Tecan. page 9 Painting a picture of gene expression throughout the mouse brain The Allen Institute in Seattle, USA, used Tecan’s GenePaint™ to compile a threedimensional map of the mouse brain. pages 10-12

Automated protein in-gel digestion for MALDI-TOF-MS Tecan’s liquid handling technology enables innovative solutions for in-gel protein digestion. pages 24-25

Work starts on the largest monoclonal antibody production facility in the Southern hemisphere Tecan and the MATF team up to create a new high throughput production system. page 5 Three-in-one platform for automated filtration, separation and plate washing Tecan’s HydroFlex™ provides increased productivity in a range of applications. page 6 Freedom EVO® Clinical 75, now available for nucleic acid testing The compact Freedom EVO Clinical has been specially configured for nucleic acid extraction and testing. page 7

Managing thousands of samples for answering ancestry questions Family Tree DNA in Texas, USA, relies on the REMP Small-Size Store™ to manage its enormous number of samples. pages 22-23

Supplying DNA to support worldwide medical research The Freedom EVO® platform handles samples that researchers worldwide rely on for their work. page 13 Automated blood pooling ensures safe PCR diagnostics Safe and reliable blood pooling with the Te-PoolSafe™ module pages 14-15

Datasweep cleans up customer complaint handling Tecan is using innovative methods to ensure customer satisfaction. page 26 Talk to Tecan...

Meet Markus Keller, head of Tecan’s global expert line and the help desk in Switzerland. page 27 Events 2007 Meet Tecan at these events before the end of the year.

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P RO D U CT N EWS

Tecan’s new QuadChamber™ is validated for use with Agilent’s 4-pack Microarrays Agilent aCGH 4-Pack Microarray

Tecan HS 4800TM with four extension units

Agilent 4-Pack Microarray multipack

Tecan has recently developed the QuadChamber for fully automated processing of four different microarrays simultaneously on one slide, using the HS Pro™ automated hybridization station. The QuadChamber was specially developed for use with Agilent’s Technologies new 4 x 44k 4-Pack Gene Expression as well as CGH Microarrays, which consist of four individual, whole-genome microarrays printed on a single glass slide. This represents the first fully automated system that can independently handle four arrays on one slide with no cross-contamination between the arrays, and is the latest development in an ongoing, global collaboration between the two companies.

The QuadChamber provides a completely sealed environment around each of the

Agilent Gene Expression 4-Pack Microarray

four arrays on Agilent’s 4-Pack slides, with independent channels for wash buffers, independent agitation mechanisms and drying for each of the four areas.

With the new chamber, the HS Pro fully automates the hybridization of the arrays from start to finish while maintaining separate compartments around each array on the slide, ensuring high quality processing with excellent reproducibility and minimal slide handling.

Agilent’s 4-Pack Microarrays concentrate sample hybridization, so provide enhanced sensitivity and allow the detection of even very low abundance transcripts. Having four arrays on each slide allows researchers to generate more genomics data per experiment while considerably reducing the required volumes of samples and reagents.

Tecan and Agilent have worked together to develop a number of protocols, including automated arraybased CGH, ensuring excellent quality data and results for customers straight away. “Agilent has worked closely with Tecan to develop our 4-Pack Microarrays and protocols for fully automated

hybridization,” said Jeff McMillan, product manager for Genomics Automation at Agilent. “The HS Pro is the only fully automated instrument that can process our 4-Pack Microarrays.”

The QuadChamber and the 4-Pack Microarrays will be presented together at a number of shows and workshops over the coming months, beginning with the Lausanne Genomics Days 2007, a symposium in Switzerland on 13-14 September, and continuing to the American Society of Human Genetics conference on 23-27 October in San Diego, USA.

Tecan’s new HS Pro QuadChamber The products described are for research use only

Tecan Journal 3/2007

PRODUCT N EWS

Enhance your crystallography with the new

CrysScreen™

2.0 software

An upgraded version of the software offers a number of user-friendly improvements in the management of protein crystallography trials.

Tecan has recently upgraded its CrysScreen protein crystallography software. The new version 2.0 runs under Tecan’s Freedom EVOware® standard software, ensuring simple and efficient crystallography process control using the Freedom EVO® liquid handling workstations. It contains a number of user-friendly improvements, including a data export function. The CrysScreen package provides easy management of protein crystallography trials, including buffer creation, set-up of experimental conditions, running of experiments, data management and analysis of results.

Watch out for further information on CrysScreen 2.0 very soon!

Work starts on the largest monoclonal antibody production facility in the Southern hemisphere Tecan and the Monash Antibody Technologies Facility (MATF) at Monash University, Australia, have announced a partnership to create one of the largest, most sophisticated monoclonal antibody production systems in the world. Plans are already underway to develop the system, which will be based around multiple, integrated liquid handling workstations and will fully automate all the stages of the production process, as well as incorporating state-of-the-art, third party devices.

The MATF is one of the only high throughput production facilities in the world offering custom-made, high-affinity monoclonal antibodies. The new system will increase its production capacity from original estimates of 500 panels of novel antibodies a year to potentially 5,000, helping to alleviate the bottleneck of antibody supply experienced by many researchers working on protein function and physiological processes. The project is partially funded by the Victorian State

Government, the Australian Stem Cell Centre and Monash University.

Tecan’s team of experienced application specialists and engineers will build the new system at the company’s headquarters in Männedorf, Switzerland, ready for installation at the MATF in April 2008.

MATF workers (l to r) Director Alan Sawyer and Deputy Director Michael Spiegel Tecan Journal 3/2007

P RO D U CT N EWS

Three-in-one platform for automated filtration, separation and plate washing Tecan’s HydroFlex™, the compact 3-in1 application platform for automated vacuum filtration, magnetic bead separation and microplate washing, provides increased productivity and reliable results by automating a range of 96-well format applications, including PCR clean-up and bead assays, ELISAs, cell assays and protein arrays. The HydroFlex automates key process steps such as

self-sealing of filtration plates, on-line vacuum control and fast dispensing of wash buffers, and offers a tunable vacuum range from -50 mbar to -850 mbar for both high and low vacuum applications.

HydroControl™ software, which has been designed to meet the FDA CFR 21 part 11 regulations. The platform can be integrated with Tecan’s Freedom EVO® liquid handling workstations or Freedom EVOlyzer® ELISA analyzer.

The HydroFlex is easily operated as a stand-alone instrument with its builtin keypad, or by using the intuitive

The HydroFlex platform for automated vacuum filtration, magnetic bead separation and microplate washing is available in various colours

Tecan Journal 3/2007

PRODUCT N EWS

Freedom EVO® Clinical 75 now available for nucleic acid testing to the Freedom EVO Clinical family has been designed and built to meet the IVDD (98/79/EC) requirements, as with the larger Freedom EVO Clinical 100, 150 and 200 platforms.

Handheld barcode scanner option

Tecan has launched the Freedom EVO Clinical 75 NAT workstation, a compact pipetting platform specially configured for extracting genomic DNA, viral DNA or RNA from clinical samples. It automates nucleic acid purification from serum or plasma, and allows flexible input from one to 24 samples; the workstation is fully compatible with the Abbott mSample Preparation System RNA and DNA kits. The extracted RNA or DNA showed excellent assay performance with Abbott RealTime® HIV, RealTime HCV and RealTime HBV viral load assays during customer evaluation. This latest addition

The instrument is equipped with a 2tip liquid handling (LiHa) arm and the Te-MagS™ magnetic bead separation module, with an adapted 24-position block allowing simultaneous pipetting of two samples; a barcode scanner can also be integrated with the system for sample identification. Tecan’s Freedom EVOware® Standard software controls the instrument and allows dedicated and open-mode scripts to be run. The integrated Sample Tracking software wizard provides complete tracking and documentation of all samples throughout the process.

Not for sale in the USA and Canada.

RealTime is a trademark of Abbott Laboratories.

Optimized layout for 24 samples

Quality and quantity control for pooling of clinical samples Tecan has recently completed its first installation of a Freedom EVO Clinical liquid handling workstation combined with the Te-PoolSafe™ option and the Pressure Monitored Pipetting (PMP) option, controlled by Freedom EVOware Sample Oriented software.

The integration of both options with the platform brings a new level of safety and quality assurance to automated blood pooling laboratories. For the first time, precise monitoring of both the dispensed volume and quality of each pipetted sample can be simultaneously performed, providing greater process security.

The Te-PoolSafe allows a fast and sensitive liquid arrival check by monitoring each pipetting step and comparing every

single dispense made with the expected volumes. The PMP tool analyzes pipetting quality by detecting faults such as clots, air aspiration, non-dispensed samples or system leakage; faults are detected by comparing recorded and real-time simulated pipetting pressure signals. The pressure simulation relies on fluidics, user-defined parameters (such as sample type and pipetting volume) and dynamic fitting of sample type and systemspecific variations (eg. in sample viscosity or system liquid properties). Tecan’s extensive prevalidation work of the two mainly used liquid classes - serum and water - allows quick and easy PMP installation and application set-up.

Typical pooling set-up with Te-PoolSafe

Not for sale in the USA and Canada.

Tecan Journal 3/2007

g lobal news

The 6th International BioForum & BioExpo Japan Proclaimed as Asia’s largest bio-event, the 6th International BioForum & BioExpo Japan was held at the Tokyo International Exhibition Center in Tokyo Bay, also known as Tokyo Big Sight, on June 20-22, 2007.

During the three-day exhibition there were almost 19,000 visitors, consisting mainly of researchers from universities, research centers and medicine/food manufacturers.

The International BioExpo attracted around 600 national and international companies, ranging from instrument and chemical manufacturers to suppliers of bioinformatics solutions and bioventures, supporting various life science research areas like pharmaceuticals, gene therapy, genomics and proteomics.

With ‘automate your cell-based assays with Tecan’ as the theme, Tecan Japan demonstrated cell-based automation solutions based on the Freedom EVO® workstation, together with other product line-ups including the latest microplate readers and washers like the Infinite™ F500 and the HydroFlex™. In the joint booth, REMP showed the Sample Safe™,

Tecan Journal 3/2007

the newly released, fully automated -80 °C sample storage and retrieval system. Tecan’s staff were so inundated with visitors to the booth, that some needed to book appointments in advance for the product specialists to be available for more detailed discussion on specific products and solutions for the customers’ application needs. A Freedom EVO 200, fully equipped with a liquid handling arm, MultiChannel Arm™ and robotic handling arm, operated continuously during the exhibition, and the Te-Flipper™ undoubtedly attracted a lot of attention from passing visitors.

The BioForum was held concurrently with the BioExpo, with a total of 250 presentations, including keynote talks from eminent life scientists. There were also research presentations by universities and public institutes in the BioAcademic Forum, technology presentations in the BioVenture Forum, and presentations about products/ technologies that support bioresearch by various exhibiting companies. Dr Titus Kretzschmar, Vice President of Research and Development at amaxa AG and Eric

Dr Titus Kretzschmar (amaxa) and Eric Willimann (Tecan) presented a seminar on automated cell culture and transfection screening

Willimann, Marketing Manager, Cell and Protein Science at Tecan, made a joint presentation entitled ‘Automated cell culture and transfection screening’ in the Exhibitors’ Product/Technology Seminar. This seminar introduced Tecan’s Freedom EVO integrated with amaxa’s Nucleofector® 96-well Shuttle® transfection system, and detailed automation solutions for high throughput screening and RNAi screening, fully automated cell culture, preparing cell lines, cell function analysis and cell-based assays. Many pharmaceutical companies participated and, even after the seminar, they were enthusiastically asking questions at the exhibition booth.

Nucleofector and 96-well Shuttle are are covered by patent and/or patent-pending rights owned by amaxa

liquid handlin g and robotics

The Tecan Integration Group Europe team (l to r) Uwe Heuzeroth, Roger Kurrat, Christoph Latzer, Hans-Peter Sattler, Isabel Patocchi, Marcel Jenny, Daniel Moser, Patrick Gut, Christoph Bosshard, Viktor Zeltner, Peter Schmidt, Petra Heerklotz, Beat Coray, Lucas Lüthy, Patrick Manz

Custom-made for you Based in Männedorf, Switzerland, Tecan Integration Group (TIG) Europe is a team of 20 highly experienced engineers and project managers, offering tailor-made automated solutions for biopharma, genomics, forensics and protein science laboratories.

Uwe Heuzeroth, head of project management, tells us more about TIG Europe. microplate carriers, adapter plates, unique parts customized for particular systems, or cappers and decappers that are specifically designed to handle different consumables or tube sealers; and can integrate a wide range of third party instruments, such as incubators, microscopes, centrifuges, washers, sealers, shakers and readers.

Patrick Gut designs an integrated system

TIG Europe provides a full project management service, from initial consultation through planning and realization to final site acceptance, mainly for customers in Europe. At TIG, we don’t simply supply the platforms and modules, but deliver them as fully customized systems, based on our Freedom EVO® platform and complex robotic systems, such as the Tecan Robotic Assay Composer (TRAC™), complete with support and information to make sure that the customer fully understands the capabilities of the system. All the systems we develop undergo thorough risk assessment and are certified to ISO standards.

As well as specializing in the large, fully automated systems, we also develop, build and integrate custommade modules, for example, special

All of TIG Europe’s project managers and engineers have a great deal of technical and software knowledge, as well as familiarity and experience in the realization of biological assays. This means that, after preliminary discussions with the customers in the very early stages, our teams can quickly determine the best ways to realize each customer’s specific requirements and achieve the required workflow.

We are organized into one engineering team plus two project management teams, and include a number of scientists with specialist knowledge in a range of subject areas, so TIG Europe is able to handle a wide variety of projects. We understand what the customers want and, with an interesting mixture of different projects that we tackle, looking for technical solutions to biological applications is really enjoyable! Recently Dr Hans-Peter Sattler was the assigned manager for the Boehringer Ingelheim

project for developing the company’s third high throughput screening system (p. 18-19). This comprehensive system was a full-on project, completely managed

by TIG Europe, and took a little over six months from order to site acceptance.

And, in the coming months, we will be working closely with our customers at the Monash Antibody Technologies Facility (MATF) to develop one of the largest, most sophisticated monoclonal antibody production systems in the world today (p. 5). Life is always exciting at TIG Europe!

Urs Zühlke sets up a robot arm

Mike Guidi and Uwe Heuzeroth, project managers at work Tecan Journal 3/2007

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A P P L I C AT I O N B I O P H A R M A

A composite image, taken from the Allen Reference Atlas, of a brain section in the coronal

plane. The left half is a Nisslstained image, and the right half is a graphic illustration delineating the brain structures.

Painting a picture

of gene expression throughout the mouse brain

Scientists at the Allen Institute for Brain Science in Seattle, USA, have recently completed the Allen Brain Atlas, a threedimensional map detailing the expression of over 21,000 genes throughout the mouse brain. This comprehensive, three-year study depended on five Tecan GenePaint™ systems for the automated in situ hybridization of gene probes in hundreds of thousands of brain sections, providing patterns of expression for every gene down to the cellular level. The atlas is freely available at www.brain-map.org.

Tecan Journal 3/2007

The Allen Institute for Brain Science was launched in 2003, with the major aim of performing innovative basic research on cellular function in the brain and the key principle to make its data and tools freely available to the public as rapidly as possible. The Allen Brain Atlas, the institute’s first project, was started in 2003 and completed in September 20061, and represented a comprehensive survey of every available, known gene in the genome. “The idea for the Allen Brain Atlas was originally defined when Institute founder and philanthropist Paul Allen convened a group of world class neuroscientists to answer the question ‘What is the one thing that could make

A P P L I C AT I O N B I O P H A R M A we had a sufficiently high throughput platform to meet our demands, and the GenePaint systems seemed to be the best available option. Another critical factor is the quality of the data, which has been sufficiently high for the scientific community to accept. The platform is also very easy to use and we have found it to be really reliable.”

Multiple assembly of slides containing brain sections into the flow-through chambers

the biggest difference in the field of brain science?’,” explained Kelly Overly, Ph.D., Research Alliance Manager at the Allen Institute.

Each gene has been individually mapped out across a series of 16 to 20 sagittal brain sections (25 µm thick) taken from fresh-frozen mouse brains (56 day-old, male C57BL/6J mice). In addition, a subset of the genes was also mapped across a series of approximately 60 coronal brain sections. The institute’s molecular biology laboratory designed and developed the probes for every gene of interest and these were automatically hybridized to the brain sections using the Tecan GenePaint systems.

“We simply place the slides onto the GenePaint systems for automated labeling and staining of the sections,” said Kelly. “We used existing ISH protocols that we adapted for the the GenePaint system.”

Each GenePaint processing cycle takes between 22.5 to 23 hours to run a batch of 192 slides so, with five systems, the researchers can process nearly 1,000 slides in parallel every day. The hybridized slides are then taken to the institute’s automated imaging facility, a series of microscopes equipped with digital cameras and driven by software programs that allow numerous slides to be loaded onto the microscope for individual imaging. The microscopes’ autofocusing algorithms can identify where each tissue section is on the slide and then take a series of images from each section at high magnification (allowing cellularlevel resolution). Each series of pictures is then stitched together to produce a high resolution image of the entire tissue section. One specific cross-section of the brain is a composite of 100 images on average, up to a high of 140 images; these composite images can be freely viewed using the on-line Allen Brain Atlas application. The atlas includes functions for zooming in or out of the images as well as the Brain Explorer visualization tool, which provides computationally reconstructed three-dimensional images of individual gene expression patterns throughout the whole brain.

The three-year project mapped the expression of over 21,000 genes using an automated in situ hybridization (ISH) platform consisting of five GenePaint systems, Tecan’s innovative platform for a variety of automated ISH, fluorescence ISH (FISH) and immunohistochemistry applications. “The automated GenePaint platform was a core part of the project,” Kelly said. “To put it in context, one of our scientists worked in a traditional academic research laboratory prior to joining the atlas project and he mapped just ten genes in five years!”

The Allen Institute originally chose GenePaint because of the previous success that Dr Gregor Eichele, one of the scientific advisory board members, had experienced with the platform.

“We scaled up the application that Dr Eichele had set up at the Baylor College of Medicine in Houston, Texas,” explained Paul Wohnoutka, Director, Atlas Operations. “We began with two Tecan GenePaint systems and, over a nine-month period, built this up to five systems. It was very important that

Assembled flow-through chamber with slide carrying brain tissue sections after in situ hybridization (ISH). The tissue sections show particular genes (purple precipitation) in specific areas of the mouse brain

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A P P L I C AT I O N B I O P H A R M A “Since its completion in 2006, we have continued to enhance the atlas with the aim of making it increasingly useful to the scientific community,” Kelly added. “Our software developers and bioinformatics teams are developing more browsing and analysis tools to allow researchers to readily find those pieces of information that are most relevant to their work, or to identify biologically relevant patterns that might only be seen with such a comprehensive collection of data. We are also continuing our annotation of the data, and all five of the GenePaint systems are still being used for ongoing projects.”

The atlas website receives approximately 10 million hits per month and a number of peer-reviewed research papers have already been published that use data from the atlas, including a study of

Tecan Journal 3/2007

human memory2 and another study that looked at glial cell differentiation and potential risk factors for multiple sclerosis3.

The Allen Institute has started to apply the GenePaint systems to additional projects. “Now that we have this great automated platform and infrastructure for high throughput ISH, we are applying it to a diversity of collaborative research projects with outside institutions, thus further helping the scientific community,” Kelly said. “We are also beginning some work on a particular part of the human brain, the neocortex, and are exploring opportunities to help advance research on brain diseases and disorders, such as autism, epilepsy and schizophrenia.”

References 1. Lein ES et al. (2007). Genome-wide atlas of gene expression in the adult mouse brain.

Nature 445: 168-176 2. Papassotiropoulos A et al. (2006). Common Kibra alleles are associated with human memory performance. Science 314: 475-478 3. Dugas JC et al. (2006). Functional genomic analysis of oligodendrocyte differentiation. J Neurosci 26: 10967-10983 The applications described here are available for research use only.

Four chamber racks containing a total of 192 flow-through chambers on a Freedom EVO 200.

Eight pipetting tips are used to pipette various reagents, buffers and wash solutions onto the slides at precisely programmed intervals, allowing the ISH procedure to be performed in an automated fashion.

LIQU I D HAN DLI NG AN D ROB OT I C S

Supplying DNA to support worldwide medical research At the Coriell Institute for Medical Research, the Coriell Cell Repositories serve the needs of the entire scientific research community by maintaining the world’s largest collection of human cell lines. Scientists there are using a Freedom EVO® 150 liquid handling workstation to prepare DNA from these cells, to distribute to research scientists investigating human disorders worldwide.

The Coriell Cell Repositories maintain a variety of cell lines representing a broad range of human diseases that are either known to have, or suspected of having, a genetic component, encompassing samples from various cancers and disorders like Prader-Willi syndrome, autism and Alzheimer’s disease. “The role of the Molecular Biology Laboratory is to prepare and provide DNA samples from these cells, for distribution to the scientific community,” explained Dr Patrick Bender, Director of the Molecular Biology Laboratory. “We have a Freedom EVO 150 workstation configured with an 8-channel liquid handling (LiHa) arm, a robotic manipulator (RoMa) arm and hotels, and an integrated REMP plate sealer which uses a variety of materials to seal 96-well plates.”

“At present, DNA quantitation is done manually, but all subsequent pipetting steps are automated. We typically prepare milligram amounts of DNA, and dispense these into microgram aliquots for shipping. We are faced with somewhat different liquid handling needs to a research laboratory, one of which is the flexibility to dispense into a variety of different vessels, such as 2

ml plastic cryovials and 96-well plates, depending on the needs of the end user.

In addition, because we dispense DNA of relatively high concentration, there are pipetting issues caused by high viscosity. The Freedom EVO performs outstandingly with water, and we were able to make a variety of measurements to establish liquid handling classes and calibration curves at various volumes for the more viscous samples we handle, using different disposable pipette tip sizes ranging from 10 µl to 1000 µl. We are pleased with the CVs, which are typically less than 8 % at 2 µl and below 5 % for volumes above 5 µl.”

“Some of the attractive features of the Freedom EVO are its flexibility and the assortment of accessories that can integrate with the instrument, such as the plate sealer. The Tecan user interface is programmable, generally by simple drag-and-drop operations. One of my colleagues attended a Tecan training course at the Research Triangle and can now easily program the instrument for a variety of laboratory processes.” Dr Bender concluded: “We have had the Freedom EVO since the beginning of 2007 and we are very happy with the platform’s performance and reliability.”

The Freedom EVO in use at Coriell

William Beggs and Susan Jones

Ron Soltesz, William Beggs, Patrick Bender, Susan Jones and Holly Bansky Tecan Journal 3/2007

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C L I N I CA L D I AGNOSTICS

Automated blood pooling ensures safe PCR diagnostics The Blood Donor Center in Halle, Germany, is using the Freedom EVO® Clinical 150 with integrated Te-PoolSafe™ module for safe and reliable automated pooling of blood for subsequent PCR-based testing, ensuring that blood products are free of infection.

The constant availability of blood for transfusions is essential for saving lives, not only of people who suffer serious accidental injuries, but also of patients undergoing surgery – a thoracic operation, for example, can require 20-30 units of blood. Blood is truly a critical commodity because, at present, there is no artificial alternative to human blood collected from blood donors. To compound this problem, blood donors, who are already in short supply, undergo a rigorous selection process, so every single donation is precious. Patients are dependent on thorough blood testing to ensure that the blood they receive is free from infections and pathogens, so it is vital that testing results are accurate and unambiguous.

Blood donor centers aim to attract potential donors and to provide blood that is as safe as current technology makes possible. Using PCR- and ELISAbased investigations in parallel, the blood donor centers confirm the results of each Tecan Journal 3/2007

The team of the laboratory from left to right: Susan Glaser, Katrin Kahle, Amelie Bolte, Heidrun Busch and Angelika Stöcker

test, while avoiding false positive results, to ensure that the patient receives ‘clean’ blood. In addition, donations undergo a set of common serological screening and blood typing tests. Many of these tests are common throughout most European countries, although some authorities demand additional blood tests.

The Blood Donor Center at the University Clinical Center in Halle, Germany, handles 100-120 blood collections each day, from which different types of blood products are prepared, and used mainly to supply the University Clinical Center hospital. The Blood Donor Center’s laboratory, led by Dipl. Biol. Angelika Stöcker, receives samples of each blood donation in barcode-labeled 13 mm

tubes, which are immediately pooled using the Freedom EVO Clinical 150 workstation. The workstation is equipped with an 8-channel liquid handling arm for four disposable and four fixed tips, 1 ml syringes, a PosID™ barcode identification device and Logic software™.

The connection to MAK-SYSTEM software allows remote communication with the workstation, eg. sending worklists to the Tecan platform from the central laboratory PC. This liquid handling platform has been upgraded by the integration of the TePoolSafe module, an automated liquid arrival check system which uses a fast and sensitive balance to weigh the pooled samples, allowing blood banks to

CLI N ICAL DIAGNOST I C S

Tecan Freedom EVO Clinical 150 workstation

monitor and evaluate the performance of their pooling application. The TePoolSafe measures the weight of every single dispense in each pool, and ensures full sample traceability by providing documented proof of performance.

When inaccurately dispensed samples are detected, the whole pool is separately re-pipetted.

The samples are processed in groups of 16 tubes. After centrifugation, 400 µl of the serum is pipetted from each of the 16 samples and pooled, after which the pools are verified using the Te-PoolSafe module. In addition, three archiving microtiter plates are prepared in case the PCR needs to be repeated, as well as aliquots of each sample for long-term storage, and stored at -30 to -40 °C.

Dispensing into the pooling tube on the Te-PoolSafe balance

the correct pool volume, whereas with the Te-PoolSafe, the 16 sample data of each pool are automatically read by the PosID barcode reader and, after pooling, a report is printed out showing the weight of every single dispense. Backed up with documented evidence, the operator can be certain that the correct volumes of every sample have been included in each pool. Unlike the manual procedures used previously, extra personnel are not

needed to watch the operator’s pooling procedure, so automation of blood pooling has yielded more walkaway time, and has freed up personnel to perform other duties.

The Te-PoolSafe option has not been cleared for use in all countries. Contact your local sales office for specific information.

Close-up view of the Te-PoolSafe balance dispensing sera into the pooling tube

PCR-based tests are performed on the pools against the human immunodeficiency virus (HIV), hepatitis B virus (HBV) and hepatitis C virus (HCV).

Any PCR that gives a positive result is repeated and, if this confirms the initial positive result, two pools of eight samples are prepared from the original 16 samples for a further PCR-based test, and then each of the eight sera from the pool that returned a positive result are individually tested. ELISA-based tests are also performed to confirm the results obtained by PCR, as well as for antibodies against human cytomegalovirus and Treponema pallidum.

Before automation of the pooling process, the operator visually inspected Tecan Journal 3/2007

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L I Q U I D H A N D L I N G A N D R O B OT I C S

Processing 15 million aliquots to support future medical research UK Biobank is using 11 Freedom EVO® liquid handling workstations to process thousands of samples each day in preparation for a long-term initiative aimed at improving the prevention, diagnosis and treatment of a wide range of serious and life-threatening illnesses, including cancer, heart diseases, diabetes, arthritis and forms of dementia.

Launched in April 2007, the UK Biobank is a major UK medical research initiative based in Manchester and set to develop an extensive resource for long-term medical studies. Over the next four years, UK Biobank will be recruiting 500,000 UK residents, aged between 40 and 69.

Detailed information about lifestyle, work, health and medical history will be collected from each volunteer, along with consent for access to the participants’ hospital and general practitioners’ medical records throughout their life.

Basic physical parameters like weight, height, blood pressure and lung function

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will also be recorded, and blood and urine samples will be taken from the volunteers.

“Over the course of 10 to 20 years, subsets of the 500,000 people will suffer from different diseases, and UK Biobank will have a package of information, together with blood and urine samples, which can be made available to research groups in various disease areas,” explained Paul Downey, Director of Operations of UK Biobank.

“Two of the key strengths of UK Biobank are the sample size and the large

amount of health information we are collecting on participants. These make UK Biobank the most detailed study of its kind ever undertaken.”

An efficient processing system is absolutely essential to handle and store so many blood and urine samples. Paul described the sheer scale of the task: “Six tubes of blood and one tube of urine are collected from each participant, and shipped overnight to our central facilities in Manchester to be processed the following day. The seven tubes are split into 28 tubes for storage so, with

L I Q U I D H A N D L I N G A N D R O B OT I C S an average daily target of 600 to 800 participants, we create roughly 20,000 1.4 ml tubes each day, and 15 million over the duration of the whole project. There were two key issues in the selection of a system to process these samples, the first of which was the reliability and throughput required to process 20,000 tubes in a day. This was critical because another 3,500 Vacutainer® collection tubes, resulting in 20,000 storage tubes, arrive each day and, as a labile substance, blood must be processed within 24 hours of collection. The other key consideration was the data audit trail, ensuring full traceability of all samples using barcodes and supported by a LIMS. After looking at several liquid handling systems, we are confident that we have chosen the system that best addresses these issues.”

The system at UK Biobank has 11 Tecan Freedom EVO liquid handling workstations integrated with various modules, such as barcode scanners and full storage hotels for tube plates, to achieve fully automatic processing of the different blood and urine tube types.

Some of the workstations are automated within laminar flow cabinets to maintain sterility of some blood samples that may be used to prepare immortalized cell lines. Once the blood or urine tubes are loaded, the entire process is automated.

The system identifies the sample tube type by its barcode, decaps the tube and aliquots the various blood or urine fractions into smaller, 2D-labeled, 96-well format storage tubes. The samples are stored at two geographically separate sites so that even if a catastrophic event at one site destroys the stored samples, the samples of any participant would not be completely lost. Two workstations have been designated for each sample type, so that there is effectively a running standby to ensure that the daily throughput can be achieved even if one platform is out of operation.

“The systems will be undergoing ISO 9001:2000 accreditation, which means that the new set-up on altered production systems needs to be validated before resuming live production, and having two of each system type allows us to do that without any downtime. Because of this strategy, combined with good reliability and efficient service from both

the local service engineer and the Tecan Integration Group (TIG), we have never been left in a situation where we can’t process samples.”

“The systems were integrated by TIG in Switzerland, and I was very impressed with their slick and professional management of the project from order to delivery. The systems were installed in stages so we have had them for between 12 and 18 months, and we have been very happy with their performance and reliability. We performed a pilot study last year to demonstrate that the project was viable, after which we gradually put the facilities and infrastructure in place before starting the project in April.”

“Currently we are ramping up the throughput and will have six or seven regional assessment centers recruiting participants around the country by the end of this year. We have just opened our third assessment center so are at around 50 % of our anticipated capacity, but I don’t see any major problem in increasing and sustaining the target daily throughput – the capacity is in place and the early signs are that everything will be fine.” Paul concluded: “I think, on balance, the project has had a really smooth start and I see it continuing that way.”

Loading sample tubes onto the Freedom EVO workstation

Barcoded tubes of blood samples

Vacutainer is a registered trademark of Beckton, Dickinson and Company.

Close-up of the tubes after centrifugation

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A P P L I CAT I O N B IOPHARM A

High throughput screening (HTS) systems at Boehringer Ingelheim The Tecan Integration Group (TIG) is currently putting together Boehringer Ingelheim’s third fully automated TRAC™-based screening system to meet the growing throughput needs of the screening laboratories at its principal research site in Biberach, Germany.

The HTS automation support team (l to r): Helga Steiner, Michael Karnath and Robert Ries

Tecan Journal 3/2007

As one of the world’s top pharmaceutical companies, Boehringer Ingelheim invests heavily in research and development, and its laboratories at Biberach are currently focusing on three therapeutic areas: diseases of the CNS, metabolic disorders and respiratory illnesses. The high throughput screening (HTS) group, led by Dr Martin J Valler, is involved in a range of essential functions associated with lead discovery, including assay adaptation, primary screening, dose response, secondary assay development and secondary screening. The HTS automation support team is led by Robert Ries. “We cover automation support for all three screening laboratories here in Biberach. We run our fully automated HTS systems 24 hours a day, five days a week,” Robert explained. “The research laboratories design the assay, and it is the day-to-day task of the four of us within the HTS group to program the automated systems to perform the assays to their specifications and required throughput. Throughput is important, because these assays are based on compound libraries from the Boehringer compound collection. There are about 950,000 compounds which we have to test for each disease target, so we require throughputs of between 50,000 and 100,000 datapoints in 24 hours, depending on the assay type. We handle many different screening projects, with different requirements, assay types and technologies, so flexibility in our automation infrastructure is essential in order to support all these demands.”

A P P L I C AT I O N B I O P H A R M A

All three TRAC systems in the HTS group are highly customized with numerous integrated devices, and have been designed to have both a very high throughput and flexibility.

Operational since 2005, the first of the systems is configured to automate 384-well microtiter plate-based high content screening (HCS) assays, and has a throughput of approximately 40 plates per 24 hours. A Thermo robot is responsible for the plate movement within the system, with a Tecan Freedom EVO® workstation and a Te-MO™ 384 pipetting device for liquid handling.

The second system has been operational since September 2006, and is designed for high throughput cell-based screening, and for biochemical assays in 384- and 1,536-well formats. Per 24 hours, it is capable of a throughput of two hundred and twenty 384-well plates, or more than sixty 1,536-well plates. In addition, this system has integrated storage devices, including cell incubators, to maintain all the assay and compound plates within the system for a 24 hour run.

by 2005 we had progressed to 1,536 as an option. We have to keep up with the very fast pace of progress and, for the smooth and efficient integration of new devices it is important for us to have a partner like TIG.” “Furthermore, Tecan has FACTS™, a very powerful flexible and modular scheduling software, which allows the integration of every device and technology you might need, including third party devices, whereas most companies are only interested in integrating their own devices. With our long experience in screening automation technology we have our own preferred technologies and devices which are not available from a single company, so it is a great advantage for us to have a partner like TIG to help us

realize the automation and integration of all these components.”

“Service is a vital aspect for us. We need a fast and competent service because our systems need to be as reliable and as stable as possible, so support with a short response time is very important.

We have support from both Germany and the TIG in Männedorf, where all the engineers for the mechanical, electrical and software development are available for us. In my experience over the last nine years, each system needs an upgrade every one or two years, and this is a neverending story. So, just in order to do the development, it is so important to have a stable partner for the future.” Robert concluded: “In this respect, TIG has been ideal for our purposes.”

The new system currently being built by Tecan in Männedorf will be configured for radioactive, biochemical and cell-based assays. A radiometric imaging reader is integrated for radioactive assays, and the concept includes all relevant precautions, such as radioactive shielding.

“TIG is perfect for our customized solutions, because we need a high level of customization in order to truly fulfil all our requirements,” continued Robert.

“Very few companies on the market have the flexibility and capability to achieve this high level of customization.

Tecan’s inherent flexibility allows us to quickly integrate new devices or replace devices, so that we can upgrade each of our systems at a future date. This is an essential advantage of the TRAC concept because we need to be able to use new technology and new devices as they become available, and particularly to accommodate miniaturization. Over the years we have rapidly miniaturized our assays; we started by using 96-well plates but by 1998 we were using 384 wells and

High throughput screening at Boehringer Ingelheim: robots at work

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A P PF TL IWA CATRIEO N B IOPHARM A SO

Clever use of

Under the seventh amendment to the EU cosmetics directive, the cosmetic industry is obliged to phase out most animal experiments for cosmetic ingredients by March 2009. Cosmital SA, a research division of Wella AG and part of Procter & Gamble, is using innovative artificial human skin models to develop and validate new methods with the goal of replacing animal testing, and relies on Tecan’s Infinite™ M200 microplate reader for many of its assays.

Dr Andreas Zeller, head of the genotoxicity laboratory at Cosmital, explained the impact of this EU directive on Wella’s Experimental Product Safety department: “Wella as a company has always avoided animal testing as much as possible and has limited this to rare cases when testing was needed to meet regulatory or safety obligations.

We do our best to be prepared for a safety assessment of new ingredients without toxicological data from animal experiments, however, finding and validating new methods before this deadline is a tremendous challenge for the cosmetic industry.”

Tecan Journal 3/2007

“With regard to detection of cytotoxicity we face many challenges, not least because one of our focus areas is hair dyes – colored chemicals that can interfere with the colorimetric assays usually associated with cytotoxicity measurement. These chemicals are often very reactive and do not wash off completely after incubating with the cells, leaving reactive residues that interfere with the assay and can give false negative or false positive values. Another problem is that the traditional endpoint assays, such as Trypan blue, resazurin or LDH release, often do not have a very broad linear range and so are not suitable for larger numbers of cells.”

“We chose the Infinite M200 reader to help us develop and measure all kinds of endpoint reactions, including cytotoxicity assays, and we also use it to quickly and simply get the spectra of the dyes we are using. Previously, we have used two basic spectrophotometers and we wanted to add fluorescence and luminescence to our capabilities. When we saw that an Infinite reader could do all these things and that there was a monochromator option available, it became clear that this would be the ideal choice. Working with all kinds of colors as we do would be completely impractical for filterbased wavelength selection and, at the same time, we have effectively replaced

APPLICATION B IOPHA R M A

three readers with just one, saving on laboratory space. However, what I like most about the Infinite M200 reader is its ease of use. Colleagues in neighboring departments use it regularly, for example, for luminometric assays. Our staff rotates around the different areas, so it is essential to have an instrument that is easy to operate. The Infinite M200 really is so easy to use that everybody knows how to operate it after just five minutes of instruction.”

At present the department is, among other things, developing a genotoxicity assay based on the reconstructed 3D skin models – a project sponsored by Colipa (the European Cosmetic, Toiletry and Perfumery Association). Dr Zeller added: “We are currently working on a Comet assay with these skin models which should allow the sensitive detection of genotoxic substances without sacrificing specificity. A significant lack of the latter is a big problem in the area of genotoxicity and leads to a huge number of so-called ‘irrelevant positives’ in in vitro testing. Detection of cytotoxicity is an important aspect of this project and the Infinite M200 helps us with this part of the protocol.”

Tecan’s Infinite M200

Cosmital genotoxicity group (l to r): Céline Procureur, Linda Corbino-Giunta, Dr Andreas Zeller

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SA M P L E M A NAG EME NT

Managing thousands of samples for answering ancestry questions The Texas-based company Family Tree DNA provides genealogy testing for people who are researching their family trees and is currently heavily involved with the Genographic Project1 mapping human migration patterns over time. The company uses PCR-based technologies to analyze DNA samples, relying on a REMP Small-Size Store™ and a Tecan Freedom EVO® liquid handling workstation to manage, process and store the hundreds of samples it receives every day.

Family Tree DNA was set up in Houston, Texas, USA, in 1999, specifically to provide DNA genealogy services on request for customers worldwide. The company analyzes DNA extracted from customers’ cheek swab samples – currently around 200 samples each day – to compare the likeness of chromosomal markers in potentially related samples, helping people to establish whether they have a common ancestor. A number of different tests are available, based on either Y chromosome testing for identifying relatedness between males, or mitochondrial (mt)DNA comparisons for establishing relatedness between females.

Family Tree DNA is also now receiving samples from volunteers participating in the Genographic Project. This five-year, worldwide study was launched in April 2005 by National Geographic and IBM, with support from the Waitt Family Foundation.

The project aims to map historical human migration patterns by collecting and analyzing DNA samples from over 100,000 people across five continents. Members of the general public are encouraged to take part and the samples are tested to determine what migratory routes the participants’ deep ancestors followed (on either the maternal or paternal side), and to which branch of the human family tree each participant belongs.

The Genographic Project has unsurprisingly led to a large increase in sample numbers for the company and the decision was quickly made to update the laboratory’s sample management system and automated processes. “I knew from previous experience that manually storing and picking hundreds of individual sample tubes can be extremely time consuming and very prone to error, so we looked for a more reliable method of cherry-picking and a more compact way to store DNA samples,” said Dr Thomas Krahn, Chief Scientific Officer at Family Tree DNA. “After investigating several sample management systems, we chose the REMP technology because

Scientists at Family Tree DNA (l to r):

Brent Manning, Astrid-Maria Krahn, Dr Connie Bormans and Dipl-Ing Thomas Krahn

Tecan Journal 3/2007

SA MPLE M ANAGEM ENNTS T COMPONA it seemed to be the most efficient. Our REMP Small-Size Store (SSS) was installed in January 2007 and we are very happy with the system; it works very well and the REMP Sample Administration Software™ is a real advantage too. We currently have around 4,000 samples in the store; its total capacity is 82,560 so we are using under five per cent capacity at the moment, but our sample numbers are increasing so quickly that within a year we expect the store to be at least three quarters full, predominantly with scientifically interesting samples and samples from customers who have reordered more markers.”

“We also chose the REMP 96 Tube Technology™ consumables,” continued Dr Krahn, “using 300 µl volume tubes because they can be capped, and also because it is vital that the tubes hold sufficient DNA for the various tests. Each customer sample yields approximately 200 µl of 5 ng/µl DNA, which is the highest concentration that we can get in a 200 µl volume.”

Family Tree DNA keeps part of every original sample in its REMP SSS in case the customer requests further tests, and also maintains a library of samples with rare alleles for control purposes.

The company currently offers about 150 different DNA tests, including tests for 12, 25, 37 or 65 markers on the Y chromosome, and tests for parts of the hypervariable regions 1 and 2 of mtDNA,

as well as full mtDNA sequencing.

“Analyzing more markers provides higher resolution data and increases the chances of identifying small mutations that can be used to reconstruct a family tree or to separate different hereditary lines,” Dr Krahn explained. “Some of our customers are involved in surname-based projects with hundreds of potential relatives, and they are keen to understand the different family lines.”

About three quarters of the samples come from customers who order straightforward analyses and do not reorder tests, and these are sent on to colleagues at a testing laboratory at the University of Arizona for processing. “Here in Houston, we focus on the specialty tests that are ordered less frequently and those that have a high degree of diversity, such as the indigenous samples sent in for the Genographic Project. All the samples to be tested are subjected to PCR, followed by various genetic analysis procedures, such as sequencing, realtime PCR or fragment length capillary electrophoresis, depending on the test required.”

Each sample usually undergoes the same DNA extraction and storage preparation procedure, so Dr Krahn and his colleagues have set up an automated platform that includes a Tecan Freedom EVO liquid handling workstation, with an integrated REMP Reatrix™ DataMatrix scanner for identifying barcodes on tubes as well

DNA templates

swabs

blood

forensic material

PCR products

Sequencing products Dye

storage in REMP SSS™ at -20°C

More than 150 different assays

CE

sequencing analysis

CE

fragment analysis

sequencing terminators

multiplex unlabeled

SNaPshot®

multiplex fl. labeled

STR capillary electrophoresis

fragment analysis

multiplex fl. labeled

Indel capillary electrophoresis

fragment analysis

PCR DNA extraction with Tecan Freedom EVO® 200

Dye terminators

Reference 1. For more information on the Genographic Project, visit www.nationalgeographic.com/ genographic Some of the referenced instrumentation is not for use in clinical diagnostics.

Electropherograms

singleplex unlabeled

+dNTP

“We are about to fully automate the entire procedure, from registering the sample tube that we receive from the customer to filling the REMP plates with purified DNA and storing them in the SSS,” Dr Krahn added. “The software for this is currently being developed and it will mean we simply register the barcode of each tube on arrival and everything will be automatically stored in the REMP database, from which we will be able to handle the orders completely independently.”

The Family Tree DNA laboratory workflow

The FTDNA laboratory work flow Customer samples

as tube racks, and a REMP Automated Capper Decapper™ (ACD96) device. The Freedom EVO performs all DNA extraction steps and purified samples are plated into REMP 96 Tube Technology tubes, placed into REMP 96 Storage Tube Racks and stored at -20 °C in the SSS. Any sample can then easily be accessed and grouped for testing as required.

PCR

Quantitative real-time PCR (TaqMan®, Plexor™)

cycle threshold analysis

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L IQ U I D H A N DLI N G AN D ROB OTICS

Automated protein in-gel digestion for MALDI-TOF-MS Olaf Jahn, Dörte Hesse, Thomas Liepold, Marina Reinelt, Hartmut D. Kratzin Proteomics Group, Max Planck Institute of Experimental Medicine, Göttingen

In-gel digestion is a major bottleneck for large scale proteome analyses and is therefore a prime candidate for automation. The Proteomics Group at the Max Planck Institute (MPI), Göttingen, in collaboration with Tecan, has developed an innovative automated solution based on Tecan liquid handling workstations.

Two-dimensional gel electrophoresis with subsequent in-gel digestion of the separated proteins followed by the analysis of those peptides using matrixassisted laser desorption ionization time-

of-flight mass spectrometry (MALDI-TOFMS) is one of the fundamental techniques of modern proteome research. A major restriction, however, is the bottleneck created by in-gel digestion and, for this reason, the Proteomics Group at MPI Göttingen, in association with Tecan, has developed a system that automates in-gel digestion, extraction of the proteolytic peptides and sample preparation for MALDI-TOF-MS, all on a single pipetting platform. Liquid handling is a major technical challenge during in-gel digestion, because very small volumes of liquid are repeatedly added and removed, readily leading to loss of sample and blockage of the pipetting needles, but the objective was to achieve the same quality of results as with the manual method while reducing hands-on time to a minimum.

Figure 1: Tecan Genesis Freedom liquid handling workstation, set up for automated in-gel digestion and sample spotting on MALDI target plates. 1, liquid handling arm with eight pipetting needles; 2, robotic manipulator arm with in-gel digestion stack (see Fig. 2a for detail); 3, cooling rack with cover for consumable reagents; 4, carrier for MALDI target (front) and downholder position for separating the in-gel digestion stack (rear); 5, cooling rack for microplates (rear) and test tubes (front); 6, carrier with supporting areas for metal lid; 7, Tecan Te-VacS vacuum separation module; 8, microplate hotel; 9, microplate incubator with temperature control.

Tecan Journal 3/2007

The system the team has developed is based on a Genesis Freedom® liquid handling workstation equipped with a liquid handling arm with eight pipetting needles, a robotic manipulator arm and a Te-VacS™ vacuum separation module (Fig. 1). Two cooling racks, with lids to prevent evaporation, ensure that the consumable reagents are kept cool, while a microplate incubator allows incubation steps at 45 °C or 50 °C. A specially designed in-gel digestion stack is made up of four components (Fig. 2a), including a 96-well protein digestion plate developed for this purpose by ABgene®/ Thermo Fisher Scientific. Each well of the digestion plate has two 250 µm holes to allow the aspiration of reagents using the Te-VacS. The protein digestion plate fits

Figure 2: In-gel digestion stack for automated in-gel digestions. a, sandwich assembly; b, stack configuration on the vacuum module

for pipetting and aspiration of reagents; c, stack configuration on the vacuum module for the elution of tryptic peptides into the collecting plate.

1, microplate cover plate with silicone mat;

LIQU I D HAN DLI NG AN D ROB OT I C S into a holder on a frame and is covered with a heavy metal plate perforated with 96 holes. A pre-perforated silicone mat on the underside of the metal plate protects the samples from evaporation and contamination and can be penetrated by the pipetting needles, while the intrinsic weight of the metal lid prevents the stack from lifting during withdrawal of the pipetting needles.

eliminates loss of sample and prevents blockage of the pipetting needles.

The peptide eluates collected are then spotted onto a MALDI target plate (Bruker Daltonics AnchorChip™ 600/384). The entire sequence of steps is controlled by flexible and fully programmable Tecan software, and the whole technical setup can also be installed on the Tecan Freedom EVO® pipetting platform.

After electrophoretic separation by SDS-PAGE and staining with Coomassie® Brilliant Blue G-250, 1.5 mm gel plugs, equivalent to 29 fmol to 34 pmol protein per gel plug, are punched out of the protein bands, inserted into the protein digestion plate and placed in the stack (Fig. 2b). The plugs are destained with ammonium bicarbonate, dehydrated with acetonitrile, dried at 50 °C and reduced with dithiothreitol (DTT). The plugs are then alkylated with iodoacetamide in the dark, washed, dehydrated and dried again, in order to ensure optimal uptake of the trypsin solution. Trypsin digestion is carried out for two hours at 45 °C and stopped by the addition of trifluoroacetic acid. Tryptic peptides are extracted by passive elution and recovered on the Te-VacS (Fig. 1, no. 7, Fig. 2c, no. 5) into a 96-well collecting plate (Fig 2c, no. 9) by contactless aspiration, which practically

At present, the new automated method can process 192 in-gel digestion samples automatically in eight hours, and validation experiments have confirmed that its efficiency is at least equivalent to the manual method (Fig. 3). The automated procedure takes an hour longer than the manual method because of the pipetting needle washing steps but, after the preparation time of about an hour, which is the same for both methods, there is no further hands-on time required. In comparison, the manual method for in-gel digestion ties up one person, almost full time, for about seven hours (Fig. 4) so the labor saving is considerable, and, by using a scheduling software, the automated throughput could be increased further without changing the technical set-up. In addition, the liquid handling arm ensures precise positioning of the samples on the

2, 96-well protein digestion plate with 250 µm holes (ABgene/Thermo Fisher Scientific); 3, holder for protein digestion plate; 4, bottom frame; 5, Tecan Te-VacS vacuum separation module; 6, pipetting needles; 7, gel plug; 8, waste; 9, 96-well collecting plate for peptide eluates.

MALDI target plate, improving the reproducibility of results1.

Overall, the method of automated sample preparation in conjunction with the MALDI-TOF-MS analysis has made routine protein identification very reliable and easy. The system described provides automated and verified high throughput proteome analyses with a high standard of significance and reproducibility, and can be implemented in principle in any laboratory. MPI is now using this method to conduct large scale proteome analyses of various biological samples, including myelin from mouse brain, frog oocytes and peroxysomes from plant leaves.

Reference 1 Olaf Jahn, Dörte Hesse, Marina Reinelt, Hartmut D. Kratzin: Technical innovations for the automated identification of gel-separated proteins by MALDI-TOF mass spectrometry. Anal.

Bioanal. Chem. (2006) 386: 92-103 Coomassie® is a registered trademark of Imperial Chemicals Industries (ICI), Ltd. ABgene® is a registered trademark of Thermo Fisher Scientific. AnchorChip™ is a trademark of Bruker Daltonics.

Figure 3: Typical results of automated in-gel digestion, comparing the automated method (black bars) with the manual method (white bars).

The charts show the data for two of the six standard proteins used, bovine serum albumin (66 kDa) and ovalbumin (54 kDa), analyzed at three different protein concentrations.

The bars represent the mean percentage sequence coverage from two independent in-gel digestions and sample preparations with twin samples (n = 4). The error bars show the standard deviation.

Figure 4: Time required for 192 in-gel digestion samples. Both the automated (A) and the manual (M) process need approximately one hour of preparation time (green). In contrast to the automated solution, this is followed by further hands-on times (red) in the case of the manual method.

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Q UA L I T Y AS SU R ANCE

Datasweep cleans up customer complaint handling A new initiative has been launched at Tecan to revolutionize the handling of customer complaints and help us to respond more quickly to our customers’ questions and suggestions.

The traditional paper-based procedures are being completely transformed into an electronic format based on Datasweep, a recently implemented software system. The new system gives worldwide access to all customer complaints and is helping to build up active dialogue between Tecan and our customers, providing a platform by which we can more easily exchange product and application related issues.

truly efficient information management, as it allows all records relating to every investigation to be stored electronically.

Once the information comes in to Datasweep, our complaint unit carries out a full investigation which requires a great deal of liaison between our production team, manufacturers and application specialists. Datasweep allows all authorized staff to access the records, wherever they are in the world, so we can all work together much more easily now and resolve each complaint more quickly and efficiently.”

The new complaint handling system has been configured with an interface between Datasweep and Tecan’s initial call management system, where Tecan help desks around the world record information of all customer calls.

Where applicable, that information is automatically transferred into Datasweep for the Quality, Customer Service and Regulatory Affairs departments to use when carrying out further investigations.

Importantly, Datasweep allows us to

“As well as being customer friendly, our new system is much more environmentally friendly!” Christie added.

“The previous method required hard copies of every document relating to each investigation to be manually stored for an indefinite period of time and all this paper is no longer required. Datasweep has so many benefits that it has been well worth all our hard work!”

Christie Richárd and Hailey Hamalainen study the Datasweep system

Tecan Journal 3/2007

look at the bigger picture, at customer feedback at product or business unit level, or even at the whole company level, allowing us to isolate any common trends and implement preventive actions as a result.

Christie Richárd, Global Compliance Director and Tecan US Quality Manager, was responsible for the implementation and validation of Datasweep, which went live in May 2006: “Datasweep provides

SE RVICES AN D CONSUM A B L E S

Talk to Tecan…

Meet Markus Keller, head of Tecan’s global liquid handling expert line and the help desk in Switzerland.

My background is in engineering and, before joining Tecan six years ago, I worked for 15 years with a number of companies specializing in technical and customer support. I head a team of four people here at Tecan Switzerland, covering the global expert line for liquid handling products, as well as the first line help desk for local Swiss customers.

Tecan has a two-stage complaint handling process with a network of local help desks all over the world.

Our people at the local help desks speak the customers’ language and, in 90 % of cases, can help the customer straightaway with their questions. The remaining 10 % of enquiries need more in-depth knowledge and are passed to the corresponding expert line, where they are addressed by technical and application specialists in that respective product area.

Whatever the enquiry, it is important to us that the customer always gets a first response from Tecan within 24 hours. All enquiries are documented and closely

The helpdesk in Switzerland, (l to r) Markus Keller and Oliver Sauter, receives calls from both local and global customers

tracked, from the initial call to the local help desk through to the expert line and including any Tecan departments involved in the process, until the issue has been satisfactorily resolved.

My team is in a special situation as we cover both levels of the complaint handling process to some extent. We deal with all sorts of enquiries, ranging from a simple request from a local customer concerning potentially any Tecan product, to a complex liquid handling problem passed to us by a colleague in another country. Every day is different!

I get great job satisfaction from working so closely with both customers and colleagues from all over the world. Some problems are not as easy to solve as others but being able to help customers and see the results straightaway makes my job so rewarding. Our customers expect excellent standards of complaint handling services from Tecan and we aim to exceed their expectations!

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EV E N TS 2 0 07

Meet Tecan at these events before the end of the year Americas International Symposium of Human Identification (ISHI)

Hollywood, CA

01 Oct 2007

American Society of Human Genetics (ASHG)

San Diego, CA

23 Oct 2007

National CODIS Conference

San Francisco, CA

29 Oct 2007

Society for Neuroscience

San Diego, CA

03 Nov 2007

American Association of Pharmaceutical Scientists

San Diego, CA

11 Nov 2007

The American Society for Cell Biology (ASCB)

Washington, DC

1 Dec 2007

China International Medical Equipment Fair

Chengdu, China

24 Oct 2007

Japanese Society for DNA Polymorphism Research

Osaka, Japan

15 Nov 2007

Biochemistry and Molecular Biology 2007

Yokohama, Japan

11 Dec 2007

The Lausanne Genomics Days 2007

Lausanne, Switzerland

13 Sep 2007

DGTI Jahreskongress

Friedrichshafen, Germany

18 Sep 2007

RNAi Europe

Barcelona, Spain

20 Sep 2007

Biotech Forum & Scanlab

Stockholm, Sweden

24 Sep 2007

ILMAC

Basel, Switzerland

25 Sep 2007

ELRIG/SBS – Drug Discovery

Nottingham, UK

01 Oct 2007

Biotechnica

Hannover, Germany

09 Oct 2007

Annual Conference of the German Society of Genetics

Jena, Germany

11 Oct 2007

Lab 07

Lillestrom, Norway

16 Oct 2007

15. Jahrestagung der Deutschen Gesellschaft für Immungenetik

München, Germany

18 Oct 2007

Medica

Düsseldorf, Germany

14 Nov 2007

Congress of the Swiss Proteomics Society

Lausanne, Switzerland

03 Dec 2007

Asia and Pacific

Europe, Middle East and Africa

Tecan Journal, Customer Magazine of Tecan Trading AG., ISSN 1660-5276 Design: OTM/London www.otmcreate.com Photography: Marc Wetli/Zürich www.wetli.com, Günter Bolzern/Zürich www.bolzern.net, Susanne Völlm/Zürich www.susannevoellm.ch Editor: kdm/UK www.kdm-communications.com Print: DAZ Druckerei Albisrieden AG/Zurich www.daz.ch Address: Tecan Switzerland AG, Marketing Communications, Seestrasse 103, CH-8708 Männedorf, Switzerland, journal@tecan.com,www.tecan.com Tecan Group Ltd. makes every effort to include accurate and up-to-date information within this publication, however, it is possible that omissions or errors might have occurred. Tecan Group Ltd. cannot, therefore, make any representations or warranties, expressed or implied, as to the accuracy or completeness of the information provided in this publication. Changes in this publication can be made at any time without notice. All mentioned trademarks are protected by law.

For technical details and detailed procedures of the specifications provided in this document please contact your Tecan representative.

This brochure may contain reference to applications and products which are not available in all markets. Please check with your local sales representative.

CrysScreen, FACTS, GenePaint, HS 4800 Pro, HydroControl, HydroFlex, Infinite, Logic software, MultiChannel Arm, PosID, QuadChamber, Te-Flipper, Te-MagS, Te-MO, Te-PoolSafe, Te-VacS and TRAC are trademarks and Freedom EVO, Freedom EVOlyzer, Freedom EVOware and Genesis Freedom are registered trademarks of Tecan Group Ltd., Männedorf, Switzerland.

Tecan is in major countries a registered trademark of Tecan Group Ltd., Männedorf, Switzerland.

Automated Capper/Decapper, Reatrix, Sample Administration Software, Sample Safe, REMP Small-Size Store and REMP Tube Technology are trademarks of REMP AG, Oberdiessbach, Switzerland. © 2007 Tecan Trading AG, Switzerland, all rights reserved.

Headquarters: Tecan Group Ltd., Seestrasse 103, CH-8708 Männedorf, Switzerland T +41 44 922 88 88 F +41 44 922 88 89 info@tecan.com www.tecan.com Tecan Austria +43 62 46 89 33 Belgium +32 15 42 13 19 China +86 10 586 95 936 Denmark +45 70 23 44 50 France +33 4 72 76 04 80 Germany +49 79 51 94 170 Italy +39 02 215 21 28 Japan +81 44 556 73 11 Netherlands +31 18 34 48 17 4 Portugal +351 21 000 82 16 Singapore +65 644 41 886 Spain +34 93 490 01 74 Sweden +46 31 75 44 000 Switzerland +41 44 922 89 22 UK +44 118 9300 300 USA +1 919 361 5200 ROW +43 62 46 89 33 REMP Switzerland +41 31 770 70 70 Japan +81 44 542 70 21 USA +1 508 429 2200

Tecan Journal 3/2007